1.Mechanism of Herbal Cake-separated Moxibustion in Improving Neuroimmune Inflammation in Rats with Chronic Fatigue Syndrome by Interfering TLR4/MyD88/NF-κB Pathway
Chuntao ZHAI ; Yawei HOU ; Linjuan SHI ; Yixiao WANG ; Wei LI ; Yuefeng TIAN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):140-149
ObjectiveTo explore the mechanism of herbal cake-separated moxibustion using the classical formula Xiaoyaosan in alleviating neuroimmune inflammatory responses in chronic fatigue syndrome (CFS) rats, based on the regulation of the toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear transcription factor-κB (NF-κB) signaling pathway. MethodsFifty SPF-grade SD rats aged 6-8 weeks were randomly divided into five groups: Normal group, model group, sham herbal cake moxibustion group, Chinese medicine intragastric administration group, and herbal cake-separated moxibustion group, with 10 rats in each group. Except for the normal group, all other groups underwent a 21-day modeling process, followed by behavioral testing. The herbal cake-separated moxibustion and sham herbal cake moxibustion groups received interventions at the Shenque (CV8), Guanyuan (CV4), Zusanli (ST36), and Qimen (LR14) acupoints. The Chinese medicine intragastric administration group was treated with a Xiaoyaosan suspension via gavage. Behavioral tests were conducted after 10 days of continuous intervention. Serum levels of interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α), as well as hippocampal levels of IL-1β, IL-6, TNF-α, and NF-κB, were detected by enzyme-linked immunosorbent assay (ELISA). Morphological changes in the hippocampus were observed using hematoxylin-eosin (HE) staining. Real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect mRNA expression levels of TLR4, MyD88, and NF-κB in the hippocampus. Western blot analysis was performed to detect the relative expression levels of TLR4, MyD88, NF-κB, and p65 proteins in the hippocampus. ResultsCompared with the normal group, the model group showed a significant decrease in upright times during the open field test (P<0.01), as well as significant reductions in total movement distance, resting time, and center region duration (P<0.01). In the tail suspension test, immobility time increased (P<0.01), and struggle times decreased (P<0.01). Serum and hippocampal levels of IL-1β, IL-6, and TNF-α, as well as hippocampal NF-κB levels and TLR4, MyD88, and NF-κB mRNA expression, were significantly elevated (P<0.01). After treatment, compared with the model group, the total movement distance and upright times in the open field test were significantly increased in all treatment groups (P<0.01), while resting time and center region duration were notably prolonged (P<0.05, P<0.01). Immobility time in the tail suspension test was significantly shortened (P<0.01), and struggle times significantly increased (P<0.05). Serum and hippocampal levels of IL-1β, IL-6, TNF-α, hippocampal NF-κB levels, and TLR4 and NF-κB mRNA expression were significantly reduced (P<0.05, P<0.01). Compared with the sham herbal cake moxibustion group, the herbal cake-separated moxibustion group showed a significant extension in center region duration during the open field test (P<0.05) and a significant increase in upright times (P<0.01). In the tail suspension test, immobility time was reduced (P<0.01), and struggle times increased (P<0.01). Serum TNF-α levels in the Chinese medicine intragastric administration group were significantly reduced (P<0.01), while serum IL-6 levels, as well as hippocampal levels of IL-1β, TNF-α, NF-κB, and TLR4, MyD88, and NF-κB mRNA expression, were significantly decreased in both the Chinese medicine intragastric administration group and the herbal cake-separated moxibustion group (P<0.05, P<0.01). Compared with the Chinese medicine intragastric administration group, the herbal cake-separated moxibustion group exhibited significantly increased upright times in the open field test (P<0.01). In the tail suspension test, immobility time was reduced (P<0.01), and struggle times increased (P<0.01). Serum IL-1β, hippocampal TNF-α levels, and TLR4, MyD88, and NF-κB mRNA expression were significantly decreased (P<0.05, P<0.01). ConclusionHerbal cake-separated moxibustion effectively improves fatigue and memory function in CFS rats, regulates neuroimmune inflammatory responses, and its mechanism may be related to the modulation of the TLR4/MyD88/NF-κB signaling pathway.
2.Development and validation of a prognostic nomogram model for patients with the lower third and abdominal oesophageal adenocarcinoma
Zhengshui XU ; Dandan LIU ; Jiantao JIANG ; Ranran KONG ; Jianzhong LI ; Yuefeng MA ; Zhenchuan MA ; Jia CHEN ; Minxia ZHU ; Shaomin LI
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2025;32(02):201-207
Objective To establish an individualized nomogram model and evaluate its efficacy to provide a possible evaluation basis for the prognosis of lower third and abdominal part of oesophageal adenocarcinoma (EAC). Methods Lower third and abdominal part of EAC patients from 2010 to 2015 were chosen from the SEER Research Plus Database (17 Regs, 2022nov sub). The patients were randomly allocated to the training cohort and the internal validation cohort with a ratio of 7∶3 using bootstrap resampling. The Cox proportional hazards regression analysis was used to determine significant contributors to overall survival (OS) in EAC patients, which would be elected to construct the nomogram prediction model. C-index, calibration curve and receiver operating characteristic (ROC) curve were performed to evaluate its efficacy. Finally, the efficacy to evaluate the OS of EAC patients was compared between the nomogram prediction model and TNM staging system. Results In total, 3945 patients with lower third and abdominal part of EAC were enrolled, including 3475 males and 470 females with a median age of 65 (57-72) years. The 2761 patients were allocated to the training cohort and the remaining 1184 patients to the internal validation cohort. In the training and the internal validation cohorts, the C-index of the nomogram model was 0.705 and 0.713, respectively. Meanwhile, the calibration curve also suggested that the nomogram model had a strong capability of predicting 1-, 3-, and 5-year OS rates of EAC patients. The nomogram also had a higher efficacy than the TNM staging system in predicting 1-, 3-, and 5-year OS rates of EAC patients. Conclusion This nomogram prediction model has a high efficiency for predicting OS in the patients with lower third and abdominal part of EAC, which is higher than that of the current TNM staging system.
3.Mechanism of immunosuppression in rats by herb-partitioned moxibustion based on transcriptome sequencing technology
Yuefeng TIAN ; Luojie XIONG ; Huifang WANG ; Chuntao ZHAI ; Wei LI
Chinese Journal of Tissue Engineering Research 2025;29(5):978-988
BACKGROUND:Immunosuppression leads to impaired body immune function and aggravates the disease.Herb-partitioned moxibustion can effectively regulate immune function and improve immunity in the body,but its regulatory mechanism has not been elucidated. OBJECTIVE:To sequence immunosuppressed model rats treated with herb-partitioned moxibustion using bioinformatics techniques based on transcriptomics and to explore the mechanisms by which it regulates immunity. METHODS:Twenty-four Sprague-Dawley rats were randomly assigned to three groups:control,model,and herb-partitioned moxibustion groups,with eight rats in each group.The model and herb-partitioned moxibustion groups were subjected to establishment of an immune suppression model by intraperitoneal injection of cyclophosphamide at a dose of 35 mg/kg for 3 consecutive days.No interventions were administered to the control and model groups after modeling.In contrast,the herb-partitioned moxibustion group received moxibustion treatment at Zhongwan,Shenque,Guanyuan,and Zusanli acupoints using a combination of moxa and herbal cakes,once a day,for 10 consecutive days,with samples being collected the day after the end of the intervention.Peripheral blood was collected from all groups of rats to measure their white blood cell count.RNA-seq was performed on the Illumina sequencing platform,and differentially expressed genes were selected for bioinformatics analysis using the GO and KEGG databases. RESULTS AND CONCLUSION:Compared with the control group,the model group exhibited a significant decrease in white blood cell count(P<0.001).RNA-seq analysis identified 3 026 differentially expressed genes between the model and control groups,with 1 565 upregulated and 1 461 downregulated.There were 535 differentially expressed genes identified between the herb-partitioned moxibustion group and the model group,with 280 upregulated and 255 downregulated.The Venn diagram analysis revealed that 159 genes were downregulated in the model group compared with the control group.However,after moxibustion with herbal cakes,these genes were upregulated.Protein-protein interaction network analysis identified 10 core targets,including Oasl,Oas2,Isg15,Herc6,Mx2,Helz2,Mx1,Syk,Hspa1a,and Ret.According to GO and KEGG analyses,moxibustion with herbal cakes regulated the body through pathways related to immune response,viruses,angiogenesis,and the autoimmune system.To conclude,there is a significant association between herbal cake-separated moxibustion intervention and immune suppression targets,including Oasl,Oas2,Isg15,Herc6,Mx2,Helz2,and Mx1.The intervention exhibits regulatory effects in the pathways related to immune responses,viral activities,and angiogenesis.
4.The Effects of Tai Chi Training on Bone Density,Bone Turnover Markers,and Heart Rate Variability in High-Risk Osteoporosis Population
Jiaming LIN ; Chao LI ; Wei ZHAO ; Jun ZHOU ; Xiaoying CHEN ; Xiangyu XI ; Haijun HE ; Baohong MI ; Yuefeng CHEN ; Weiheng CHEN
Journal of Traditional Chinese Medicine 2025;66(15):1566-1571
ObjectiveTo explore the effects of the Tai Chi training on bone density, bone turnover markers, and heart rate variability for people with high-risk osteoporosis, and to provide evidence for the prevention of osteoporosis at early stage. MethodsSixty-six cases of people with high risk of osteoporosis were included, and they were divided into 33 cases each in the intervention group and the control group using the random number table method. The control group received osteoporosis health education three times a week, and the intervention group received Tai Chi training under the guidance of a trainer three times a week for 40 mins each time on the basis of the control group, and both groups were intervened for 12 weeks. Dual-energy X-ray absorptiometry was used to measure the bone density of L1~L4 vertebrae, bilateral femoral necks and bilateral total hips in the two groups before and after the intervention; enzyme-linked immunosorbent assay was used to determine bone turnover markers before and after the intervention, including pro-collagen type Ⅰ pro-amino-terminal prepropyl peptide (P1NP) and β-collagen type Ⅰ cross-linking carboxy-terminal peptide (β-CTX). Seven cases with good compliance in the intervention group were selected. After wearing the heart rate sensor, they successively performed Tai Chi training and walking activities recommended by the guideline for 20 mins each, and the heart rate variability (HRV) during exercise was collected, including time-domain indexes such as standard deviation of normal sinus intervals (SDNN), root-mean-square of the difference between adjacent RR intervals (RMSSD), frequency-domain metrics such as low-frequency power (LF), high-frequency power (HF), and low-frequency/high-frequency power ratio (LF/HF), as well as nonlinear metrics such as approximate entropy (ApEn), sample entropy (SampEn). ResultsFinally, 63 cases were included in the outcome analysis, including 30 cases in the intervention group and 33 cases in the control group. After the intervention, the differences of L1~L4 vertebrae, bone density of bilateral femoral neck and bilateral total hip in the intervention group were not statistically significant when compared with those before intervention (P>0.05), while the bone density of all parts of the control group decreased significantly compared with that before intervention (P<0.05), and the difference in the bone density of the L1~L4 vertebrae, bilateral femoral neck, and the right total hip before and after the intervention of the intervention group was smaller than that of the control group (P<0.05). The differences in P1NP and β-CTX between groups before and after intervention was not statistically significant (P>0.05). Compared with walking exercise, LF decreased, HF increased and LF/HF decreased during Tai Chi exercise (P<0.05); the time domain indexes and non-linear indexes between groups had no statistically significant difference (P>0.05). ConclusionTai Chi exercise can maintain lumbar, hip, and femoral bone density and improve sympathetic/parasympathetic balance in people at high risk for osteoporosis, but cannot significantly improve bone turnover markers.
5.Research on Material Basis of Homologous and Heterogeneous Effect of Aurantii Fructus Immaturus and Aurantii Fructus Based on Total Statistical Moment Analysis and Molecular Connectivity Index
Xiao YUAN ; Yuefeng WU ; Shixiong LI ; Youzhi LIU ; Zhiqin YU ; Fuyuan HE ; Jin ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(3):161-168
ObjectiveTo investigate the material basis of homologous and heterogeneous effect of Aurantii Fructus Immaturus(AFI) and Aurantii Fructus(AF) based on the total statistical moment analysis and molecular connectivity index(MCI). MethodRelevant literature at home and abroad and Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP) were consulted to establish the chemical composition database of AFI and AF, and set up their fingerprints by ultra-high performance liquid chromatography(UPLC), and the total statistical moments and similarity parameters of the fingerprint were calculated. According to MCI, all components of AFI and AF were divided into different component groups, the average values of 0-8th order(0χ-8χ) MCI of the common component groups of AFI and AF were calculated. ResultThe values of total zero-order moment(AUCT) of AFI and AF were (10.57±2.45)×106, (5.09±0.89)×106 μV·s, the values of total first-order moment(MCRTT) were (11.57±1.58), (12.10±1.29) min, the values of total second-order moments(VCRTT) were(24.49±2.30), (26.49±2.54) min2, respectively. It showed that qualitative and quantitative parameters of AFI and AF were significantly different. The components with high similarity such as neohesperidin, hesperidin and narirutin were screened as the common potential pharmacodynamic components of AFI and AF. The non-common components of AFI, such as alysifolinone and imperatorin, and the non-common components of AF, such as neoeriocitrin and isosakuranin, with high similarity were screened out as potential heterogeneous components of AFI and AF. The composition groups of AFI and AF were classified into six categories, and the similarities between the composition groups of AFI and AF and the total constituents were 0.872-0.979 and 0.918-0.997, the average values of 0χ-8χ MCI of alkaloids in AFI and AF were 3.65 and 3.14, the average values of 0χ-8χ MCI of flavonoids were 8.47 and 8.47, the average values of 0χ-8χ MCI of volatile oils were 2.71 and 3.48, respectively. It showed that there were some differences in MCI of chemical constituents(groups) between AFI and AF. ConclusionThe chemical constituents(groups) of AFI and AF not only differ in content and species, but also in structural characteristics and structure-activity relationship, which can provide a basis for further explaining the scientific connotation of homologous and heterogeneous effect of AFI and AF.
6.Real-World Study on the Clinical Efficacy of Different Medication Regimens of Wangbi Tablet (尪痹片) in the Treatment of Knee Osteoarthritis
Kuayue ZHANG ; Chao LI ; Zhuoyun WU ; Yawei DONG ; Zelu ZHENG ; Yuzhi LIU ; Jun ZHOU ; Jiaming LIN ; Yuefeng CHEN ; Baohong MI ; Weiheng CHEN
Journal of Traditional Chinese Medicine 2024;65(22):2316-2325
ObjectiveTo investigate the differences in clinical efficacy of different medication regimens of Wangbi Tablets (尪痹片) for knee osteoarthritis (KOA) in a real-world setting, providing a basis for rational clinical use of Wangbi Tablets. MethodsA prospective registry study was conducted, involving 2,999 KOA patients registered in 30 hospitals nationwide from January 26th, 2019, to December 17th, 2021. Based on the use of Wangbi Tablets during the observation period, patients were divided into a monotherapy group (1,507 cases) and a combination therapy group (1,492 cases), and the combination group can be further divided into Wangbi Tablets plus Chinese medicine (CM), Wangbi Tablets plus western medicine (WM), and Wangbi Tablets plus Chinese and western medicine (CM+WM) subgroups. The baseline data of patients in the monotherapy group and the combination group were compared, including age, gender, body weight, medication time, clinical stage, K-L grade, and others. Efficacy indicators included the Visual Analog Scale (VAS) score, Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score, and EuroQol five-dimensional (EQ-5D) health index, which were evaluated before and after 4-, 8- and 12-week treatment, and the difference before and after treatment was calculated after 4, 8 and 12 weeks of treatment. The difference between the baseline and 12 weeks of treatment of all the above indicators was used as the dependent variables, and gender, age, body mass index (BMI), course of disease, K-L grade, and clinical stage were used as independent variables, when multiple linear regression was taken to explore the influencing factors of the efficacy. At the same time, the occurrence of major symptoms (including morning stiffness, joint swelling, soreness of waist and knees, fear of wind, and fear of cold) was counted, and the disappearance of symptoms at each time point was counted after 4, 8, and 12 weeks of treatment. ResultsAt baseline, there were no statistically significant differences in gender and age distribution between the monotherapy and combination therapy groups (P>0.05); the proportion of patients in the acute stage and recovery stage was higher in the monotherapy group than in the combination therapy group, while the proportion in the remission stage was lower (P<0.05); the VAS score was higher in the monotherapy group, and the EQ-5D index was lower (P<0.01), with no statistically significant difference in total WOMAC score between the two groups (P>0.05). Compared to those measured before treatment and at previous timepoint, the VAS score and WOMAC total score significantly decreased in both groups, while EQ-5D value increased (P<0.05). The difference in VAS score between baseline and after 12-week treatment was higher in the monotherapy group than the combination group, while the differences in WOMAC total score and EQ-5D value between baseline and after 4-, 8- and 12-week treatment were higher in the combination group (P<0.05). Multiple linear regression showed that VAS score before treatment had greatest impact on pain improvement (P<0.01), and compared to Wangbi Tablets monotherapy, the combination of Wangbi tablets with WM or CM had larger associations with pain improvement (P<0.05); and Wangbi Tablets had better efficacy when the course of treatment was >28 days (P<0.01). Wangbi Tablets plus WM had a better effect on improving the overall function of the knee joint than Wangbi Tablets alone (P<0.01); and the efficacy of Wangbi Tablets with a course of treatment >28 days was better (P<0.05). The improvement of quality of life of patients in the attack and remission stages was more obvious than that in the recovery stage (P<0.01); Wangbi Tablets plus WM or CM had a better effect on improving quality of life than Wangbi Tablets alone (P<0.05). Before treatment, the proportion of patients with morning stiffness, soreness of waist and knees, fear of wind and chills in the monotherapy group was higher than that in the combination group (P<0.01). The proportion of main symptoms in both groups decreased after 4, 8 and 12 weeks of treatment (P<0.05). After 4 weeks of treatment, the disappearance rate of each main symptom in the combination group was higher than that in the monotherapy group, and after 12 weeks of treatment, the disappearance rate of fear of wind in the monotherapy group was higher than that in the combination group, while the disappearance rate of joint swelling and soreness of waist and knees was lower (P<0.05). ConclusionWangbi Tablets, whether used alone or in combination with other medications, is effective throughout the course of KOA, with greater benefits in improving joint function and quality of life during the acute and remission stages compared to the recovery stage. Combination therapy had a faster onset of effect, but began to converge with monotherapy after 8 weeks. The best efficacy was observed with the combination of Wangbi Tablets with WM, followed by combination with CM.
7.Influence of Pregnancy Medication on Maternal Pregnancy Outcomes and Follow-up Study
Yun HONG ; Xianhui CAI ; Lin LI ; Lu LI ; Yuefeng RAO
Chinese Journal of Modern Applied Pharmacy 2024;41(2):242-247
OBJECTIVE
To explore the current status and safety of medication in pregnant patients, and explore the working mode and data accumulation of the pregnancy medication consultation clinic.
METHODS
All pregnant women who received consultations at the Pregnancy and Lactation Medication Consultation Clinic of the First Affiliated Hospital of Zhejiang University School of Medicine from October 2021 to December 2022 were selected as the study subjects. Their names, ages, contact information, last menstrual period, and medication use during pregnancy were recorded, and the medication profile and safety of the study population were evaluated. Organized the intervention measures of clinical pharmacists and conducted follow-up studies on their pregnancy outcomes.
RESULTS
A total of 179 consultation pregnant women were included, and 80% of the patients in the study population used drugs in the unknown pregnancy state, with 3 drugs per capita. The most common reason for drug use was upper respiratory tract infection. The three types of drugs with the highest use rate were traditional Chinese patent medicines, antibacterial drugs, antipyretic and analgesic drugs. The usage rate of B-grade drugs in the study population was the highest, accounting for 51.3%; 8.4% of patients used X-grade medication. One hundred and fifty-one cases were followed up, with a loss of follow-up rate of 15.6%. The recommended acceptance rate of drug risk assessment given by pharmacists for patients was 95.4%. There were 98.7% of patients believe that pregnancy counseling clinics were helpful and could effectively alleviate anxiety.
CONCLUSION
Pregnancy medication consultation clinics can provide patients with scientific and personalized medication guidance and popular science education, and have significant effects in alleviating anxiety and depression in pregnant women.
8.BMSC-Exo alleviates depression-like behaviors in mice by inducing hippocampal energy metabolism and microangiogenesis
Weijia HU ; Lili CAI ; Xiaolan ZHU ; Yifeng LUO ; Zhihong CAO ; Yuefeng LI
Chinese Journal of Neuromedicine 2024;23(1):2-9
Objective:To investigate the effect of bone mesenchymal stem cells derived exosomes (BMSC-Exo) on improving hippocampal microangiogenesis, energy metabolism, and behaviors in depression mouse models.Methods:(1) Mouse bone marrow mesenchymal stem cells were isolated and cultured to extract BMSC-Exo; BMSC-Exo morphology was observed by transmission electron microscopy, BMSC-Exo particle diameter ranges were determined by Zetaview analyzer, and expressions of CD9 and CD63 in BMSC-Exo were detected by Western blotting. (2) Depression models were established in 2 mice by chronic unforeseeable mild stress (CUMS); 24 h after stereotaxic injection of phosphate buffer solution (PBS) or DiR labeled BMSC-Exo, BMSC-Exo uptake was detected by in vivo imaging system. (3) Thirty-six mice were randomly divided into control group, model group and BMSC-Exo group ( n=12); CUMS was used to establish depression models in the latter 2 groups; brain stereotaxic injection of 1 μL BMSC-Exo was given to mice in the BMSC-Exo group after modeling, and same amount of PBS was given to the control group; behaviors were observed by forced swimming test (FST), tail suspension test (TST) and open field test (OFT); hippocampal microvascular length and number were detected by alkaline phosphatase staining; energy metabolism in the hippocampus was detected by micro positron emission tomography/computed tomography (mPET/CT); glucose transporter 1 (GLUT1) expression in the hippocampus was detected by Western blotting. Results:(1) BMSC-Exo had a typical disk-like vesicle-like structure with particle size of (100.5±1.4) nm; Western blotting confirmed that CD9 and CD63 expressed in BMSC-Exo. (2) In vivo imaging showed no fluorescence in the brain and liver after PBS injection, but obvious local fluorescence after BMSC-Exo injection. (3) Compared with the control group, the model group and BMSC-Exo group had significantly longer rest time in FST and TST and shorter movement distance and time in the central region of OFT ( P<0.05); compared with the model group, BMSC-Exo group had significantly shorter rest time in FST and TST and longer movement distance and time in the central region of OFT ( P<0.05). Compared with the control group, the model group and BMSC-Exo group had significantly decreased standard uptake value (SUV) of regions of interest, microvascular length and number, and GLUT1 expression in the hippocampus ( P<0.05); compared with the model group, the BMSC-Exo group had significantly higher SUV, microvascular length and number, and GLUT1 expression in the hippocampus ( P<0.05). Positive correlations were noted between hippocampal microvascular length and SUV and between microvascular number and SUV in the 3 groups ( r=0.540, P<0.001; r=0.600, P<0.001). Conclusion:BMSC-Exo could promote microangiogenesis energy metabolism in the hippocampus to improve depression-like behaviors in depression mouse models.
9.Biocontrol Effect and Culture Condition Optimization of Trichoderma citrinoviride Strain Isolated from Rhizosphere of Atractylodes lancea
Kuo LI ; Sheng WANG ; Hongyang WANG ; Zheng PENG ; Xiuzhi GUO ; Tielin WANG ; Yuefeng WANG ; Lanping GUO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(15):192-199
ObjectiveRoot rot is one of the most serious diseases in the cultivation and production of Atractylodes lancea. Trichoderma spp. are effective in the biocontrol of root rot without causing environmental pollution. This study aims to isolate and study a Trichoderma strain capable preventing and controlling root rot from the rhizosphere of A. lancea and to solve the problem of disease prevention and control in the planting and production of A. lancea. MethodTrichoderma T2204 was isolated by the dilution-coating method and identified by ITS sequencing. The inhibitory activities of T2204 and its volatiles against two pathogenic fungal strains were examined by dual-culture and co-culture experiments. The biocontrol potential of T2204 on root rot of A. lancea and the effect of T2204 on the accumulation of medicinal compounds in the rhizosphere of A. lancea were investigated by pot experiments and GC-MS, respectively. In addition, the optimal medium, photoperiod, temperature, pH, and carbon and nitrogen sources for the culture of T2204 were explored. ResultThe Trichoderma isolate T2204 was identified as T. citrinoviride and had direct inhibitory effects on two highly pathogenic strains causing root rot. In the dual-culture experiments with the two pathogenic strains, T2204 showcased the inhibition rates of 77.90% and 76.80%, respectively. In the co-culture experiments with the two pathogenic strains, the volatile organic compounds produced by T2204 showed the inhibition rates of 57.11% and 81.11%, respectively. The pot experiments showed that the survival rate of A. lancea seedlings infected by root rot reached 100% after inoculation with T2204 and was only 50% in the case without inoculation of T2204. After 150 days of cultivation, the dry weight and atractylodin content of the rhizome of A. lancea plants treated with T2204 increased by 32% (P<0.05) and 11%, respectively, compared with the untreated group. The optimal conditions for the growth of T2204 were PDA or PSA medium, photoperiod of 12 h dark/12 h light, 25-30 °C, pH 5-6, carbon sources of glucose, D-fructose, soluble starch, and maltose, and the nitrogen sources of ammonium sulfate and ammonium dihydrogen phosphate. The optimal conditions for the sporulation of T2204 were PSA or CMA medium, photoperiod of 12 h dark/12 h light, 20-30 °C, pH 8, carbon source of sucrose, and nitrogen source of sodium nitrate. ConclusionT2204 could improve the growth and root rot resistance of A. lancea and promote the accumulation of medicinal compounds. The findings laid a foundation for the industrialized production and application of T2204 in the production of A. lancea in the future.
10.Exploration of the mechanism of micro ribonucleic acid-3197 in diabetic reti-nopathy based on nuclear factor κB signaling pathway
Shasha HAN ; Yuefeng LI ; Xinmeng XU
Recent Advances in Ophthalmology 2024;44(3):188-192
Objective To explore the mechanism of micro ribonucleic acid(miR)-3197 in diabetic retinopathy(DR)on the basis of the nuclear factor κB(NF-κB)signaling pathway.Methods A total of 47 DR patients admitted to Heng-shui People's Hospital from January 2021 to December 2021 were selected as the DR group,and 47 healthy individuals in the same period were collected as the control group.Their information in gender,age,fasting blood glucose(FBG),fast-ing insulin(FINS),triglycerides(TG),total cholesterol(TC)and miR-3197 were compared.The correlation between miR-3197 in DR patients and laboratory data was analyzed,and the receiver operating characteristic(ROC)curve of miR-3197 for DR diagnosis was drawn.The human retinal microvascular endothelial cells(hRMECs)were cultured in vitro and treated with 5.5 mmol·L-1 glucose[low glucose(NG)group]and 30 mmol·L-1 glucose[high glucose(HG)group],respectively.After transfecting with anti-miR-NC and anti-miR-3197,the cells were treated with 30 mmol·L-1 glucose(HG+anti-miR-NC group and HG+anti-miR-3197 group).Real-time fluorescence quantitative PCR was used to detect the relative expression level of miR-3197,flow cytometry was used to detect the apoptosis rate of hRMECs,enzyme-linked im-munosorbent assay was used for detecting tumor necrosis factor-a(TNF-a)and interleukin-6(IL-6),and Western blot was adopted to detect the expressions of aspartic protease 3 containing cysteine(cleaved caspase-3)protein,Bax protein and NF-κB signaling pathway-related proteins[phospho-NF-KB p65(p-p65),p65,phospho-NF-KB inhibited protein(p-IκBα),and NF-κB inhibited protein(IκBα)].Results The levels of FBG,FINS,TC and TG in the DR group were higher than those in the control group,and the differences were statistically significant(all P<0.001).The relative expression level of miR-3197 in the peripheral blood of patients in the DR group(2.76±0.67)was higher than that of the control group(1.03±0.34),and the difference was statistically significant(P<0.05).The miR-3197 level of patients in the DR group was positively correlated with FBG,FINS,TC and TG levels(r=0.672,0.587,0.511 and 0.423;all P<0.05).The ROC curve graph showed that the area under the curve was 0.919,with sensitivity and specificity of 85.11%and 89.36%,respectively.Compared with the NG group,the HG group showed a significant increase in cell apoptosis rate and the pro-tein expressions of cleaved caspase-3,Bax,TNF-a,IL-6,p-IκBa and p-p65(all P<0.05);compared with the HG+anti-miR-NC group,the HG+anti-miR-3197 group showed a significant decrease in cell apoptosis rate and the protein expres-sions of cleaved caspase-3,Bax,TNF-a,IL-6,p-IκBa and p-p65(allP<0.05).Conclusion The miR-3197 is highly ex-pressed in the peripheral blood of DR patients and high glucose-induced hRMECs.Down-regulation of miR-3197 can allevi-ate high glucose-induced hRMEC apoptosis and inflammatory injury,and its mechanism of action may be related to the inhi-bition of the NF-κB signaling pathway.


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